Oleanolic acid(OA)is a pentacyclic triterpenoid chemical component that exists in natural plants with a molecular formula of C30H48O3 and a molecular weight at 456.71 g·mol-1.OA is widespread in traditional Chine...Oleanolic acid(OA)is a pentacyclic triterpenoid chemical component that exists in natural plants with a molecular formula of C30H48O3 and a molecular weight at 456.71 g·mol-1.OA is widespread in traditional Chinese herbal medicine(Ligustri Lucidi Fructus,Achyranthis Bidentate Radix,Red Sage)and berries(blueberries,grapes).In recent years,because of the extensive pharmacological effects of OA,its advantages in disease treatment have become increasingly prominent and gradually attracted the attention of pharmaceutical researchers.OA has effective therapeutic effects on a series of chronic diseases such as inflammation,cancer,diabetes,and cardiovascular diseases through multiple signaling pathways and various targets.Especially in cancers,such as colorectal cancer,liver cancer,gastric cancer,lung cancer,breast cancer and other malignancies,OA presents substantial efficacy.However,its poor aqueous solubility,needy bioavailability,and unsatisfactory pharmacological activity excessively restrict its clinical application.More importantly,the improper utilization of OA can cause adverse reactions,toxic effects and even damage to organs in some specific situations.With the discovery of various pharmacological effects,the complex action mechanisms of OA,the continuous progress in structural modification of OA,as well as the synthesis of OA derivatives,its application is expanding gradually.Among numerous studies,there is a clear indication that OA and its derivatives,if fully developed,may provide an alternative and cheaper treatment for a variety of chronic diseases.However,the specific molecular mechanisms of OA and its derivatives as an alternative therapy and supplementary therapy for cancer,diabetes,cardiovascular disease and other chronic diseases remain to be clarified.Therefore,it is necessary to further study the pharmacokinetics,pharmacological activity,specific targets and related mechanisms of OA to lay a solid foundation for drug development and the application of OA in clinical settings.展开更多
OBJECTIVE To identify the inhibitory effect of ursolic acid on the colorectal cancer HCT116 cells in vitro and in vivo,and to explore the underlying mechanism.METHODS The smoothened(SMO)gene-silenced human colorectal ...OBJECTIVE To identify the inhibitory effect of ursolic acid on the colorectal cancer HCT116 cells in vitro and in vivo,and to explore the underlying mechanism.METHODS The smoothened(SMO)gene-silenced human colorectal cancer HCT116^(hSMO)cell line was established by transfection with the lentivirus carrying SMO shRNA.The cytotoxic effect of ursolic acid on HCT116^(hSMO)cells was determined by MTT assay.The effect of ursolic acid on the migration of HCT116^(hSMO)cells was studied by wound healing assay.The effect of ursolic acid on apoptosis of HCT116^(hSMO)cells was explored by Hoechst33342/PI double staining and flow cytometry.The effects of ursolic acid on the expressions of apoptotic marker gene Bcl-2,Bax,caspase-3 and caspase-9 were measured by real-time quantitative RT-PCR(RT-qPCR)and Western blotting(WB)analysis.RT-qPCR and WB were used to examine the relationship between GLI1,c-Myc expression and PI3K/Akt pathway to further investigate the mechanism of GLI1 activation in HCT116^(hSMO)cells.The effects of ursolic acid on the expressions of GLI1,p-Akt,Akt,c-Myc,SHH and SUFU of noncanonical Hedgehog pathway were evaluated by RT-qPCR and WB assays.Xenograft nude mouse model bearing HCT116^(hSMO)cells was established and intraperitoneally treated with ursolic acid to investigate the effect on tumor growth in vivo.The body weight and tumor size of mice were assessed regularly every 2 d.The effect of ursolic acid on the apoptosis of tumor tissue was determined by TUNEL assay.The expressions of Bcl-2,Bax,GLI1,p-Akt,Akt,c-Myc,SHH,SUFU mRNA and proteins were measured by RT-qPCR and WB.The levels of Bcl-2,Bax,GLI1,p-Akt,c-Myc and SHH proteins in tumor tissues were also evaluated by immunohistochemistry.RESULTS Ursolic acid significantly inhibited the growth and migration of HCT116^(hSMO)cells in vitro,compared with the control(P<0.05).Meanwhile,ursolic acid also induced apoptosis of HCT116^(hSMO)cells in vitro(P<0.05).Furthermore,SC79(Akt activator)enhanced the expressions of p-Akt,GLI1 and c-Myc,which could be abolished by ursolic acid,and the effect was equal to Akt inhibitor LY294002.The expressions of Bcl-2,GLI1,p-Akt,c-Myc,SHH mRNA and proteins were reduced by ursolic acid,while the levels of Bax and SUFU were increased.Ursolic acid could inhibit the growth and induce the apoptosis of colorectal cancer xenograft in vivo.Similarly,lower levels of Bcl-2,GLI1,p-Akt,c-Myc and SHH,and higher expression of Bax and SUFU were noted in ursolic acid-treated mice.CONCLUSION Ursolic acid can inhibit the growth and induce apoptosis of HCT116^(hSMO)cells both in vitro and in vivo.And the mechanism is related to the suppression of PI3K/Akt-mediated noncanonical Hedgehog signaling pathway.展开更多
OBJECTIVE To investigate the pharmacological effect of ursolic acid(UA)on colitis-associated colorectal cancer(CAC)and its underlying mechanism based on the Wnt signaling pathway.METHODS The CAC model in mice was esta...OBJECTIVE To investigate the pharmacological effect of ursolic acid(UA)on colitis-associated colorectal cancer(CAC)and its underlying mechanism based on the Wnt signaling pathway.METHODS The CAC model in mice was established by azoxymethane(AOM)combined and dextran sulfate sodium salt(DSS),accompanied by treatment with various dosages of UA and concomitant appraisal of body weight,stool and physical state of the mice.After the sacrifice of the mice,the tumor and length of the colorectum were measured,followed by retrieval of the liver,spleen,thymus and tumor tissue for downstream assays.The levels of inflammatory factors interleukin-6(IL-6),IL^(-1)βand C-reactive protein(CRP)in the tumor and serum were examined by enzyme-linked immunosorbent assay(ELISA).The pathological changes of colorectal tissues were observed by HE staining.The levels in tumors of Wnt/β-catenin signaling pathway-related proteins Wnt4,GSK-3β,β-catenin,TCF4,LEF1,c-Myc,cyclin D1 and apoptosis-related protein Bcl-2 were assayed by immunohistochemistry(IHC).The mRNA expressions of Wnt4,GSK-3β,β-catenin,TCF4,LEF1,c-Myc,cyclin D1,Bcl-2,Bax,caspase-9 and caspase-3 in tumors were detected by real-time quantitative RT-PCR(RT-qPCR).The protein levels of Wnt4,GSK-3β,β-catenin,TCF4,LEF1,c-Myc,cyclin D1,phospho-β-catenin,phospho-GSK-3β,Bcl-2 and Bax in tumors were probed by analyzed by Western blotting(WB).Also,RNA-seq was employed to assess the gut microbiota in the mice.RESULTS UA significantly ameliorated the symptoms of AOM/DSS-induced mouse CAC,evidenced by improved physical state,body weight,survival rate,colorectal length,the mass of liver,thymus,spleen,and decreased CAC load and colorectal mass.UA attenuated the levels of IL-6,IL^(-1)βand CRP in the mouse serum and colorectal tumor in a dose-dependent manner.HE staining showed that UA lessened carcinogenesis in the colorectum,with lower infiltration of lymphocytes,versus the control.IHC indicated that UA mitigated the expression of Wnt4,β-catenin,TCF4,LEF1,c-Myc,cyclin D1,Bcl-2,and promoted the GSK-3βexpression,compared with the control.Furthermore,UA diminished the mRNA expressions of Wnt4,β-catenin,TCF4,LEF1,c-Myc,cyclin D1,Bcl-2,and heightened the mRNA levels of GSK-3β,caspase-3,capase-9 and Bax in CAC.The results of mRNA expressions were verified by WB analysis,which revealed that UA impeded the protein expression of Wnt4,β-catenin,c-Myc,cyclin D1,Bcl-2,TCF4,LEF1,and elevated the protein levels of GSK-3βand Bax,phospho-β-catenin in mouse CAC.In addition,UA substantially ameliorated the gut microbiota to store the metabolic function in the mice with CAC.CONCLUSION Ursolic acid may protect against CAC,potentially by downregulation of Wnt/β-catenin signaling pathway activity and restoration of gut microbiota.展开更多
OBJECTIVE To investigate the inhibition and mechanism of berberine on human colorectal cancer HCT116 cells through canonical Hedgehog signaling pathway.METHODS The effect of berberine on cell morphology was observed b...OBJECTIVE To investigate the inhibition and mechanism of berberine on human colorectal cancer HCT116 cells through canonical Hedgehog signaling pathway.METHODS The effect of berberine on cell morphology was observed by microscopy.MTT colorimetric assay,cell scratch experiment,colony formation assay and Hoechest/PI staining were utilized to detect the activities of berberine on cell viability,cell migration and cell apoptosis.Flow cytometry was applied to examine the cell apoptosis.The effects of berberine on caspase-3 and caspase-9 were detected by caspase activity detection kit.The expressions of Hedgehog signaling pathway-related proteins SHH,GLI1,PTCH1,SMO,SUFU,apoptosis-related proteins Bax and Bcl-2 as well as cell cycle-related proteins cyclin D1 were detected by Western blotting.Additionally,quantitative real time RT-PCR was employed to assess the mRNA expression levels of Hedgehog signaling pathway-related genes SHH,GLI1,PTCH1,SMO,SUFU,apoptosis-related genes Bax and Bcl-2 as well as cell cycle-related genes cyclin D1.RESULTS Berberine sharply altered the morphology of human colorectal cancer HCT116 cells,demonstrated by that migration ability of HCT116 cells was reduced significantly and the nuclei were densely stained.Berberine could induce apoptosis in a dose-dependent manner.The activities of caspase-3 and caspase-9 were increased prominently.The expression levels of Hedgehog signaling pathway-related protein SUFU and apoptosis-related protein Bax were augmented substantially.The expression levels of Hedgehog signaling pathway-related proteins SHH,GLI1,PTCH1,SMO,apoptosis-related protein Bcl-2 as well as cell cycle-related genes cyclin D1 were markedly lessened.Besides,the mRNA expression levels of Hedgehog signaling pathway-related gene SUFU and apoptosis-related gene Bax were augmented substantially.The mRNA expression levels of Hedgehog signaling pathway-related genes SHH,GLI1,PTCH1,SMO,apoptosis-related gene Bcl-2 as well as cell cycle-related gene cyclin D1 were markedly lessened.CONCLUSION Berberine,which is the main component of coptidis rhizoma,can remarkably restrain the growth and proliferation,promote apoptosis of human colorectal cancer cells HCT116,and the underlying mechanism may be involved in suppressing the activity of the Hedgehog signaling pathway.展开更多
OBJECTIVE To investigate the therapeutic effect of scutellarin on colitis-associated cancer(CAC)and its underlying mechanism based on Wnt/β-catenin signaling pathway.METHODS The mouse model of CAC was established by ...OBJECTIVE To investigate the therapeutic effect of scutellarin on colitis-associated cancer(CAC)and its underlying mechanism based on Wnt/β-catenin signaling pathway.METHODS The mouse model of CAC was established by azomethane oxide(AOM)and sodium dextran sulfate(DSS),followed by scutellarin treatment,with recording the body weight,diarrhea and hematochezia.After sacrificing the mice,the colorectal length and colorectal tumor were assessed.The levels of pro-inflammatory factors TNF-αand IL-6 in mice′s sera were measured by the enzyme-linked immunosorbent assay(ELISA).The colorectal lesions were appraised by hematoxylin and eosin(H&E)staining.Theβ-catenin level in CAC tissues was probed by immunofluorescent analysis.The apoptosis-related genes Bax and Bcl-2,and Wnt signaling pathway-related genesβ-catenin,GSK-3β,TCF4,c-Myc and cyclin D1 were detected by real-time quantitative RT-PCR(RT-qPCR).Finally,Western blotting analysis(WB)was employed to examine the expressions of the apoptosis and Wnt signaling pathway-related proteins.RESULTS Scutellarin significantly improved AOM/DSS-caused weight loss,colorectal length shortening,and tumor growth in mice(P<0.01).Meanwhile,colorectal lesions could be substantially alleviated by scutellarin.ELISA results showed that the levels of pro-inflammatory factors TNF-αand IL-6 were drastically lessened(P<0.01).Scutellarin also sharply inhibited the nuclear translocation ofβ-catenin,as evidenced by the reduction in the nuclear level ofβ-catenin protein.In addition,scutellarin attenuated the mRNA expression of Wnt signaling pathway-relatedβ-catenin,TCF4,c-Myc and cyclin D1,whereas it heightened GSK-3βmRNA level.These results were consolidated by WB analysis,which indicated that scutellarin could mitigate the protein levels of phospho-GSK-3β,β-catenin,TCF4,c-Myc and cyclin D1,with the increase in GSK-3βprotein in CAC tissue.Moreover,scutellarin could induce the apoptosis of CAC,demonstrated by enhanced expression of Bax and diminished expression of Bcl-2 in both mRNA and protein levels.CONCLUSION Scutellarin may ameliorate colitis-associated colorectal cancer by weakening Wnt/β-catenin signaling cascade.展开更多
OBJECTIVE To investigate the effect of scutellarin on the apoptosis of human colorectal cancer cells via the Hippo signaling pathway in vitro.METHODS MTT colorimetric method was used to detect the influence of scutell...OBJECTIVE To investigate the effect of scutellarin on the apoptosis of human colorectal cancer cells via the Hippo signaling pathway in vitro.METHODS MTT colorimetric method was used to detect the influence of scutellarin on the survival rate of HCT116 cells.And the effect of scutellarin at various concentrations on cell morphology was observed by microscopy.Cell scratch experiment was used to detect the influence of scutellarin on the migration of HCT116 cells.Hoechst33342/PI double staining method was used to detect the effect of scutellarin on the apoptosis of HCT116 cells.Western blotting method was used to assess the action of scutellarin on the expressions of Hippo signaling pathway-related proteins Mst1,Lats1,YAP1,p-YAP(Ser127),TAZ,and its downstream effector proteins c-Myc and cyclin D1,as well as apoptosis-related proteins Bcl-2 and Bax in HCT116 cells.RESULTS Scutellarin significantly affected the morphology of HCT116 cells and reduced the survival rate of HCT116 cells.Hoechst33342/PI double staining showed that scutellarin effectively induced the apoptosis of HCT116 cells.Western blotting analysis showed that the expression levels of Hippo signaling pathway-related proteins Mst1,Lats1,YAP1,TAZ and its downstream effector proteins c-Myc,cyclin D1 were down-regulated in a concentration-dependent manner by scutellarin,and the expression of p-YAP(ser127)was up-regulated.Moreover,scutellarin substantially lessened the expression level of apoptosis-related protein Bcl-2,and promoted the protein level of Bax.CONCLUSION Scutellarin may inhibit the proliferation and migration of HCT116 cells,while induce its apoptosis,potentially by activation of Hippo signaling pathway.展开更多
Pulsatilla chinensis is a widely used traditional Chinese herb,which contains 56 types of chemical constituents,mainly including triterpenoid saponins,organic acids,coumarins and lignans.The largest portion of the ing...Pulsatilla chinensis is a widely used traditional Chinese herb,which contains 56 types of chemical constituents,mainly including triterpenoid saponins,organic acids,coumarins and lignans.The largest portion of the ingredients in Pulsatilla chinensis is the family of triterpenoid saponins,in which anemoside B4 is the major effective compound and indexing component.The main components of Pulsatilla chinensis can metabolize into a vast array of active products in vivo,which play vital roles in its biological activity.Mounting evidence reveals that Pulsatilla chinensis exerts a wide range of therapeutic activities,such as anti-cancer,immunoregulation,anti-inflammation and anti-schistosome,with fewer adverse reactions,via various signaling pathways and multiple targets.It was documented that the active ingredient of Pulsatilla chinensis can lessen the drug resistance and synergize the effects of other natural products including paclitaxel,as well as ameliorate the clinical efficacy of chemical drugs,such as adriamycin.However,Pulsatilla chinensis was also reported to be possibly the main cause of hemolysis and chronic liver injury.The efforts should be made to deeply investigate the pharmacological actions and underlying mechanisms of Pulsatilla chinensis,with a focus on the anti-cancer efficacy,and develop new drugs based on the components of Pulsatilla chinensis for future utilization in the clinical setting.展开更多
基金National Natural Science Foundation of China(81573813,81173598)Sichuan Provincial Admin⁃istration of Traditional Chinese Medicine of China(2021MS447)+1 种基金Excellent Talent Program of Chengdu University of Tra⁃ditional Chinese Medicine of China(YXRC2019002,ZRYY1917)and Open Research Fund of the State Key Laboratory of Southwestern Chinese Medicine Resources of China(2020XSGG006)。
文摘Oleanolic acid(OA)is a pentacyclic triterpenoid chemical component that exists in natural plants with a molecular formula of C30H48O3 and a molecular weight at 456.71 g·mol-1.OA is widespread in traditional Chinese herbal medicine(Ligustri Lucidi Fructus,Achyranthis Bidentate Radix,Red Sage)and berries(blueberries,grapes).In recent years,because of the extensive pharmacological effects of OA,its advantages in disease treatment have become increasingly prominent and gradually attracted the attention of pharmaceutical researchers.OA has effective therapeutic effects on a series of chronic diseases such as inflammation,cancer,diabetes,and cardiovascular diseases through multiple signaling pathways and various targets.Especially in cancers,such as colorectal cancer,liver cancer,gastric cancer,lung cancer,breast cancer and other malignancies,OA presents substantial efficacy.However,its poor aqueous solubility,needy bioavailability,and unsatisfactory pharmacological activity excessively restrict its clinical application.More importantly,the improper utilization of OA can cause adverse reactions,toxic effects and even damage to organs in some specific situations.With the discovery of various pharmacological effects,the complex action mechanisms of OA,the continuous progress in structural modification of OA,as well as the synthesis of OA derivatives,its application is expanding gradually.Among numerous studies,there is a clear indication that OA and its derivatives,if fully developed,may provide an alternative and cheaper treatment for a variety of chronic diseases.However,the specific molecular mechanisms of OA and its derivatives as an alternative therapy and supplementary therapy for cancer,diabetes,cardiovascular disease and other chronic diseases remain to be clarified.Therefore,it is necessary to further study the pharmacokinetics,pharmacological activity,specific targets and related mechanisms of OA to lay a solid foundation for drug development and the application of OA in clinical settings.
基金National Natural Science Foundation of China(81573813,81173598)Sichuan Provincial Admin⁃istration of Traditional Chinese Medicine of China(2021MS447)+1 种基金Excellent Talent Program of Chengdu University of Tra⁃ditional Chinese Medicine of China(YXRC2019002,ZRYY1917)and Open Research Fund of the State Key Laboratory of Southwestern Chinese Medicine Resources of China(2020XSGG006)。
文摘OBJECTIVE To identify the inhibitory effect of ursolic acid on the colorectal cancer HCT116 cells in vitro and in vivo,and to explore the underlying mechanism.METHODS The smoothened(SMO)gene-silenced human colorectal cancer HCT116^(hSMO)cell line was established by transfection with the lentivirus carrying SMO shRNA.The cytotoxic effect of ursolic acid on HCT116^(hSMO)cells was determined by MTT assay.The effect of ursolic acid on the migration of HCT116^(hSMO)cells was studied by wound healing assay.The effect of ursolic acid on apoptosis of HCT116^(hSMO)cells was explored by Hoechst33342/PI double staining and flow cytometry.The effects of ursolic acid on the expressions of apoptotic marker gene Bcl-2,Bax,caspase-3 and caspase-9 were measured by real-time quantitative RT-PCR(RT-qPCR)and Western blotting(WB)analysis.RT-qPCR and WB were used to examine the relationship between GLI1,c-Myc expression and PI3K/Akt pathway to further investigate the mechanism of GLI1 activation in HCT116^(hSMO)cells.The effects of ursolic acid on the expressions of GLI1,p-Akt,Akt,c-Myc,SHH and SUFU of noncanonical Hedgehog pathway were evaluated by RT-qPCR and WB assays.Xenograft nude mouse model bearing HCT116^(hSMO)cells was established and intraperitoneally treated with ursolic acid to investigate the effect on tumor growth in vivo.The body weight and tumor size of mice were assessed regularly every 2 d.The effect of ursolic acid on the apoptosis of tumor tissue was determined by TUNEL assay.The expressions of Bcl-2,Bax,GLI1,p-Akt,Akt,c-Myc,SHH,SUFU mRNA and proteins were measured by RT-qPCR and WB.The levels of Bcl-2,Bax,GLI1,p-Akt,c-Myc and SHH proteins in tumor tissues were also evaluated by immunohistochemistry.RESULTS Ursolic acid significantly inhibited the growth and migration of HCT116^(hSMO)cells in vitro,compared with the control(P<0.05).Meanwhile,ursolic acid also induced apoptosis of HCT116^(hSMO)cells in vitro(P<0.05).Furthermore,SC79(Akt activator)enhanced the expressions of p-Akt,GLI1 and c-Myc,which could be abolished by ursolic acid,and the effect was equal to Akt inhibitor LY294002.The expressions of Bcl-2,GLI1,p-Akt,c-Myc,SHH mRNA and proteins were reduced by ursolic acid,while the levels of Bax and SUFU were increased.Ursolic acid could inhibit the growth and induce the apoptosis of colorectal cancer xenograft in vivo.Similarly,lower levels of Bcl-2,GLI1,p-Akt,c-Myc and SHH,and higher expression of Bax and SUFU were noted in ursolic acid-treated mice.CONCLUSION Ursolic acid can inhibit the growth and induce apoptosis of HCT116^(hSMO)cells both in vitro and in vivo.And the mechanism is related to the suppression of PI3K/Akt-mediated noncanonical Hedgehog signaling pathway.
基金National Natural Science Foundation of China(81573813,81173598)Sichuan Provincial Admin⁃istration of Traditional Chinese Medicine of China(2021MS447)+1 种基金Excellent Talent Program of Chengdu University of Tra⁃ditional Chinese Medicine of China(YXRC2019002,ZRYY1917)Open Research Fund of the State Key Laboratory of Southwestern Chinese Medicine Resources of China(2020XSGG006)。
文摘OBJECTIVE To investigate the pharmacological effect of ursolic acid(UA)on colitis-associated colorectal cancer(CAC)and its underlying mechanism based on the Wnt signaling pathway.METHODS The CAC model in mice was established by azoxymethane(AOM)combined and dextran sulfate sodium salt(DSS),accompanied by treatment with various dosages of UA and concomitant appraisal of body weight,stool and physical state of the mice.After the sacrifice of the mice,the tumor and length of the colorectum were measured,followed by retrieval of the liver,spleen,thymus and tumor tissue for downstream assays.The levels of inflammatory factors interleukin-6(IL-6),IL^(-1)βand C-reactive protein(CRP)in the tumor and serum were examined by enzyme-linked immunosorbent assay(ELISA).The pathological changes of colorectal tissues were observed by HE staining.The levels in tumors of Wnt/β-catenin signaling pathway-related proteins Wnt4,GSK-3β,β-catenin,TCF4,LEF1,c-Myc,cyclin D1 and apoptosis-related protein Bcl-2 were assayed by immunohistochemistry(IHC).The mRNA expressions of Wnt4,GSK-3β,β-catenin,TCF4,LEF1,c-Myc,cyclin D1,Bcl-2,Bax,caspase-9 and caspase-3 in tumors were detected by real-time quantitative RT-PCR(RT-qPCR).The protein levels of Wnt4,GSK-3β,β-catenin,TCF4,LEF1,c-Myc,cyclin D1,phospho-β-catenin,phospho-GSK-3β,Bcl-2 and Bax in tumors were probed by analyzed by Western blotting(WB).Also,RNA-seq was employed to assess the gut microbiota in the mice.RESULTS UA significantly ameliorated the symptoms of AOM/DSS-induced mouse CAC,evidenced by improved physical state,body weight,survival rate,colorectal length,the mass of liver,thymus,spleen,and decreased CAC load and colorectal mass.UA attenuated the levels of IL-6,IL^(-1)βand CRP in the mouse serum and colorectal tumor in a dose-dependent manner.HE staining showed that UA lessened carcinogenesis in the colorectum,with lower infiltration of lymphocytes,versus the control.IHC indicated that UA mitigated the expression of Wnt4,β-catenin,TCF4,LEF1,c-Myc,cyclin D1,Bcl-2,and promoted the GSK-3βexpression,compared with the control.Furthermore,UA diminished the mRNA expressions of Wnt4,β-catenin,TCF4,LEF1,c-Myc,cyclin D1,Bcl-2,and heightened the mRNA levels of GSK-3β,caspase-3,capase-9 and Bax in CAC.The results of mRNA expressions were verified by WB analysis,which revealed that UA impeded the protein expression of Wnt4,β-catenin,c-Myc,cyclin D1,Bcl-2,TCF4,LEF1,and elevated the protein levels of GSK-3βand Bax,phospho-β-catenin in mouse CAC.In addition,UA substantially ameliorated the gut microbiota to store the metabolic function in the mice with CAC.CONCLUSION Ursolic acid may protect against CAC,potentially by downregulation of Wnt/β-catenin signaling pathway activity and restoration of gut microbiota.
基金National Natural Science Foundation of China(81573813,81173598)Sichuan Provincial Admin⁃istration of Traditional Chinese Medicine of China(2021MS447)+1 种基金Excellent Talent Program of Chengdu University of Tra⁃ditional Chinese Medicine of China(YXRC2019002,ZRYY1917)and Open Research Fund of State Key Laboratory of Southwestern Chinese Medicine Resources of China(2020XSGG006)。
文摘OBJECTIVE To investigate the inhibition and mechanism of berberine on human colorectal cancer HCT116 cells through canonical Hedgehog signaling pathway.METHODS The effect of berberine on cell morphology was observed by microscopy.MTT colorimetric assay,cell scratch experiment,colony formation assay and Hoechest/PI staining were utilized to detect the activities of berberine on cell viability,cell migration and cell apoptosis.Flow cytometry was applied to examine the cell apoptosis.The effects of berberine on caspase-3 and caspase-9 were detected by caspase activity detection kit.The expressions of Hedgehog signaling pathway-related proteins SHH,GLI1,PTCH1,SMO,SUFU,apoptosis-related proteins Bax and Bcl-2 as well as cell cycle-related proteins cyclin D1 were detected by Western blotting.Additionally,quantitative real time RT-PCR was employed to assess the mRNA expression levels of Hedgehog signaling pathway-related genes SHH,GLI1,PTCH1,SMO,SUFU,apoptosis-related genes Bax and Bcl-2 as well as cell cycle-related genes cyclin D1.RESULTS Berberine sharply altered the morphology of human colorectal cancer HCT116 cells,demonstrated by that migration ability of HCT116 cells was reduced significantly and the nuclei were densely stained.Berberine could induce apoptosis in a dose-dependent manner.The activities of caspase-3 and caspase-9 were increased prominently.The expression levels of Hedgehog signaling pathway-related protein SUFU and apoptosis-related protein Bax were augmented substantially.The expression levels of Hedgehog signaling pathway-related proteins SHH,GLI1,PTCH1,SMO,apoptosis-related protein Bcl-2 as well as cell cycle-related genes cyclin D1 were markedly lessened.Besides,the mRNA expression levels of Hedgehog signaling pathway-related gene SUFU and apoptosis-related gene Bax were augmented substantially.The mRNA expression levels of Hedgehog signaling pathway-related genes SHH,GLI1,PTCH1,SMO,apoptosis-related gene Bcl-2 as well as cell cycle-related gene cyclin D1 were markedly lessened.CONCLUSION Berberine,which is the main component of coptidis rhizoma,can remarkably restrain the growth and proliferation,promote apoptosis of human colorectal cancer cells HCT116,and the underlying mechanism may be involved in suppressing the activity of the Hedgehog signaling pathway.
基金National Natural Science Foundation of China(81573813,81173598)Sichuan Provincial Admin⁃istration of Traditional Chinese Medicine of China(2021MS447)+1 种基金the Excellent Talent Program of Chengdu University of Traditional Chinese Medicine of China(YXRC2019002,ZRYY1917)Open Research Fund of the State Key Labora⁃tory of Southwestern Chinese Medicine Resources of China(2020XSGG006)。
文摘OBJECTIVE To investigate the therapeutic effect of scutellarin on colitis-associated cancer(CAC)and its underlying mechanism based on Wnt/β-catenin signaling pathway.METHODS The mouse model of CAC was established by azomethane oxide(AOM)and sodium dextran sulfate(DSS),followed by scutellarin treatment,with recording the body weight,diarrhea and hematochezia.After sacrificing the mice,the colorectal length and colorectal tumor were assessed.The levels of pro-inflammatory factors TNF-αand IL-6 in mice′s sera were measured by the enzyme-linked immunosorbent assay(ELISA).The colorectal lesions were appraised by hematoxylin and eosin(H&E)staining.Theβ-catenin level in CAC tissues was probed by immunofluorescent analysis.The apoptosis-related genes Bax and Bcl-2,and Wnt signaling pathway-related genesβ-catenin,GSK-3β,TCF4,c-Myc and cyclin D1 were detected by real-time quantitative RT-PCR(RT-qPCR).Finally,Western blotting analysis(WB)was employed to examine the expressions of the apoptosis and Wnt signaling pathway-related proteins.RESULTS Scutellarin significantly improved AOM/DSS-caused weight loss,colorectal length shortening,and tumor growth in mice(P<0.01).Meanwhile,colorectal lesions could be substantially alleviated by scutellarin.ELISA results showed that the levels of pro-inflammatory factors TNF-αand IL-6 were drastically lessened(P<0.01).Scutellarin also sharply inhibited the nuclear translocation ofβ-catenin,as evidenced by the reduction in the nuclear level ofβ-catenin protein.In addition,scutellarin attenuated the mRNA expression of Wnt signaling pathway-relatedβ-catenin,TCF4,c-Myc and cyclin D1,whereas it heightened GSK-3βmRNA level.These results were consolidated by WB analysis,which indicated that scutellarin could mitigate the protein levels of phospho-GSK-3β,β-catenin,TCF4,c-Myc and cyclin D1,with the increase in GSK-3βprotein in CAC tissue.Moreover,scutellarin could induce the apoptosis of CAC,demonstrated by enhanced expression of Bax and diminished expression of Bcl-2 in both mRNA and protein levels.CONCLUSION Scutellarin may ameliorate colitis-associated colorectal cancer by weakening Wnt/β-catenin signaling cascade.
基金National Natural Science Foundation of China(81573813,81173598)Sichuan Provincial Admin⁃istration of Traditional Chinese Medicine of China(2021MS447)+1 种基金Excellent Talent Program of Chengdu University of Tra⁃ditional Chinese Medicine of China(YXRC2019002,ZRYY1917)and Open Research Fund of the State Key Laboratory of Southwestern Chinese Medicine Resources of China(2020XSGG006)。
文摘OBJECTIVE To investigate the effect of scutellarin on the apoptosis of human colorectal cancer cells via the Hippo signaling pathway in vitro.METHODS MTT colorimetric method was used to detect the influence of scutellarin on the survival rate of HCT116 cells.And the effect of scutellarin at various concentrations on cell morphology was observed by microscopy.Cell scratch experiment was used to detect the influence of scutellarin on the migration of HCT116 cells.Hoechst33342/PI double staining method was used to detect the effect of scutellarin on the apoptosis of HCT116 cells.Western blotting method was used to assess the action of scutellarin on the expressions of Hippo signaling pathway-related proteins Mst1,Lats1,YAP1,p-YAP(Ser127),TAZ,and its downstream effector proteins c-Myc and cyclin D1,as well as apoptosis-related proteins Bcl-2 and Bax in HCT116 cells.RESULTS Scutellarin significantly affected the morphology of HCT116 cells and reduced the survival rate of HCT116 cells.Hoechst33342/PI double staining showed that scutellarin effectively induced the apoptosis of HCT116 cells.Western blotting analysis showed that the expression levels of Hippo signaling pathway-related proteins Mst1,Lats1,YAP1,TAZ and its downstream effector proteins c-Myc,cyclin D1 were down-regulated in a concentration-dependent manner by scutellarin,and the expression of p-YAP(ser127)was up-regulated.Moreover,scutellarin substantially lessened the expression level of apoptosis-related protein Bcl-2,and promoted the protein level of Bax.CONCLUSION Scutellarin may inhibit the proliferation and migration of HCT116 cells,while induce its apoptosis,potentially by activation of Hippo signaling pathway.
基金National Natural Science Foundation of China(8157381381173598)+3 种基金Sichuan Provincial Admin⁃istration of Traditional Chinese Medicine of China(2021MS447)Excellent Talent Program of Chengdu University of Tra⁃ditional Chinese Medicine of China(YXRC2019002ZRYY1917)and Open Research Fund of the State Key Laboratory of Southwestern Chinese Medicine Resources of China(2020XSGG006)。
文摘Pulsatilla chinensis is a widely used traditional Chinese herb,which contains 56 types of chemical constituents,mainly including triterpenoid saponins,organic acids,coumarins and lignans.The largest portion of the ingredients in Pulsatilla chinensis is the family of triterpenoid saponins,in which anemoside B4 is the major effective compound and indexing component.The main components of Pulsatilla chinensis can metabolize into a vast array of active products in vivo,which play vital roles in its biological activity.Mounting evidence reveals that Pulsatilla chinensis exerts a wide range of therapeutic activities,such as anti-cancer,immunoregulation,anti-inflammation and anti-schistosome,with fewer adverse reactions,via various signaling pathways and multiple targets.It was documented that the active ingredient of Pulsatilla chinensis can lessen the drug resistance and synergize the effects of other natural products including paclitaxel,as well as ameliorate the clinical efficacy of chemical drugs,such as adriamycin.However,Pulsatilla chinensis was also reported to be possibly the main cause of hemolysis and chronic liver injury.The efforts should be made to deeply investigate the pharmacological actions and underlying mechanisms of Pulsatilla chinensis,with a focus on the anti-cancer efficacy,and develop new drugs based on the components of Pulsatilla chinensis for future utilization in the clinical setting.
